Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Experiment Videos

The erasable Western blot.

S H Kaufmann, C M Ewing, J H Shaper

    Analytical Biochemistry
    |February 15, 1987
    PubMed
    Summary
    This summary is machine-generated.

    Researchers developed a novel method to remove antibodies from nitrocellulose blots, preserving immobilized polypeptides. This technique allows for blot reuse, enhancing western blot efficiency and reducing experimental costs.

    Related Concept Videos

    You might also read

    Related Articles

    Articles linked to this work by shared authors, journal, and citation graph.

    Sort by
    Same author

    Erasable Western blots.

    Methods in molecular biology (Clifton, N.J.)·2012
    Same author

    Erasable Western blots.

    Methods in molecular biology (Clifton, N.J.)·2012
    Same author

    Identification of novel CHD1-associated collaborative alterations of genomic structure and functional assessment of CHD1 in prostate cancer.

    Oncogene·2011
    Same author

    Cyclooxygenase-2 is up-regulated in proliferative inflammatory atrophy of the prostate, but not in prostate carcinoma.

    Cancer research·2001
    Same author

    Multiple antibodies to titin immunoreact with AHNAK and localize to the mitotic spindle machinery.

    Cell motility and the cytoskeleton·2001
    Same author

    A novel human cell culture model for the study of familial prostate cancer.

    Cancer research·2001
    Same journal

    Lysozyme assay using a rationally designed GN4G2 substrate with coupled β-glucosidase reaction.

    Analytical biochemistry·2026
    Same journal

    The long run: A tribute to Arthur Joseph Lawrence Cooper.

    Analytical biochemistry·2026
    Same journal

    Evaluation of a method for affinity measurement using solution equilibrium titration with magnetic beads.

    Analytical biochemistry·2026
    Same journal

    Metabolomics approach using UHPLC/QE-MS for the mechanism of He Xue Ming Mu tablets on non-proliferative diabetic retinopathy.

    Analytical biochemistry·2026
    Same journal

    UniRES-GO: Unified residue-level early fusion of sequence and predicted structure for protein function prediction.

    Analytical biochemistry·2026
    Same journal

    IgG detection by enzyme-linked mass spectrometric assay versus color, fluorescent, ECL in buffer and serum.

    Analytical biochemistry·2026
    See all related articles

    Area of Science:

    • Biochemistry
    • Molecular Biology
    • Immunology

    Background:

    • Western blotting is a crucial technique for detecting specific proteins.
    • Antibody binding to nitrocellulose membranes can be irreversible, limiting blot reuse.
    • Existing methods for antibody removal may damage immobilized protein targets.

    Purpose of the Study:

    • To develop a method for efficient antibody removal from nitrocellulose blots.
    • To preserve the integrity of immobilized polypeptides for subsequent detection.
    • To enable the reuse of blots, thereby reducing experimental costs and time.

    Main Methods:

    • Polypeptides were separated using polyacrylamide gel electrophoresis (PAGE) with sodium dodecyl sulfate (SDS).
    • Proteins were transferred to nitrocellulose membranes, and nonspecific sites were blocked with nonfat dried milk.

    Related Experiment Videos

  • Blots were incubated with primary and radiolabeled secondary antibodies, followed by washing.
  • Antibodies were removed by incubating blots at 70°C in 2% SDS with 100 mM beta-mercaptoethanol.

    • Main Results:

    • The developed method successfully removed primary and secondary antibodies from nitrocellulose blots.
    • Immobilized polypeptides remained preserved on the blots after antibody removal.
    • Blots were reused for subsequent detection without compromising signal integrity.
    • A modified method also allowed for the reuse of nylon membranes.

    Conclusions:

    • This novel method provides an effective way to strip and reuse blots, increasing experimental efficiency.
    • The preservation of polypeptide integrity allows for multiple rounds of probing on the same blot.
    • The technique is applicable to both nitrocellulose and nylon membranes, offering versatility in western blotting protocols.